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High contents of both docosahexaenoic and arachidonic acids in milk of women consuming fish from lake Kitangiri (Tanzania): targets for infant formulae close to our ancient diet?

Current recommendations for arachidonic (AA) and docosahexaenoic (DHA) acids in infant formulae are based on milk of Western mothers. Validity may be questioned in view of the profound dietary changes in the past 100 years, as opposed to our slowly adapting genome. Hominin evolution occurred in the proximity of East-African freshwater lakes and rivers and early homo sapiens had higher intakes of AA and DHA from a predominantly lacustrine-based diet. In search of milk AA and DHA contents of our African ancestors, we investigated the milk of 29 lactating women living in Doromoni near lake Kitangiri (Tanzania). They consumed sunflower oil-fried local fish as only animal lipid sources, maize and local vegetables. AA and DHA contents of Doromoni milk may be close to that of early homo sapiens, because of the similarity of their life-long consumption of East-African lacustrine-based foods. Human milk fatty acid relationships from our historical worldwide database and the literature revealed that disparities between the Doromoni diet and the presumed ancient diet (i.e. higher carbohydrate and linoleic acid intakes) are unlikely to affect milk AA and DHA contents. Doromoni milk had high contents of AA (median 0.70 mol%), DHA (0.75) and eicosapentaenoic acid (EPA, 0.17), and low AA/DHA ratios (median 0.91; 0.55-2.61). This tracks down to consumption of fish with high AA and DHA contents, and AA/EPA ratios. We conclude that the milk AA, DHA and EPA contents of Doromoni women might provide us with clues to optimize infant formulae and perhaps the milk of Western women.

Adolescent↗

Determination of niacin in infant formula and wheat flour by anion-exchange liquid chromatography with solid-phase extraction cleanup.

Niacin content must be included on food labels of infant formula products and bakery products containing enriched flour. Liquid chromatographic (LC) determination of niacin in complex food matrixes is complicated by the presence of endogenous compounds that absorb at the commonly used wave-length of 260 nm. Also, the presence of particulate matter in the standard sulfuric acid extraction procedure results in reduced life of LC columns and precolumns. A simple, rapid, solid-phase extraction (SPE) procedure for separation and cleanup of niacin from a complex food matrix digest has been developed. By using a vacuum manifold with the SPE column system, multiple samples can be processed quickly and efficiently for LC analysis, compared with gravimetric column cleanup. Sulfuric acid sample digest is passed over an aromatic sulfonic acid cation-exchange (ArSCX-SPE) or a sulfonated Florisil SPE column. Niacin is eluted with 0.25M sodium acetate-acetic acid, pH 5.6 buffer in vacuo. LC chromatograms of the resulting eluate are free of interference from other components absorbing at 260 nm at the retention time of niacin. Validation of the method was obtained from agreement of analytical results on available reference materials. For both SPE methods, values for niacin in SRM 1846 Infant Formula (milk-based powder) were within uncertainty ranges of the certified value. Use of several calibration procedures (the LC computer program, a peak area response graphic standard curve, or the method of standard additions) with both SPE procedures resulted in niacin values for 3 RM-Wheat Flours (not certified for niacin) in agreement (90-105%) with their respective values reported in the literature. Several commercial wheat flours showed a broad 260 nm interference, resulting in high niacin values. Niacin recoveries from spiked soy-based liquid infant formulas ranged from 95-107% with the ArSCX-SPE column. Calibration curves of niacin were linear up to 400 micrograms/mL, with a detection limit of 0.2 microgram/mL.

Chromatography, Ion Exchange↗

Vitamin D in infant formula and enteral products by liquid chromatography: collaborative study.

Results from a collaborative study of a new liquid chromatographic (LC) method for determination of vitamin D in infant formulas and enteral products are presented. Each of 15 laboratories was provided with 11 blind duplicate samples covering a range of approximately 200-500 International Units/quart (normal dilution), a system suitability sample, and the U.S. Pharmacopeia ergo- and cholecalciferol standards. Product types included liquid and powder forms of milk (whey and casein), soy, and hydrolyzed protein-based infant formulas and enteral products. The method includes a single liquid-liquid extraction following saponification, solid-phase extraction, and then concentration by evaporation. An isocratic, nonaqueous, chromatographic system with reversed-phase, zero endcapped C18 column, and UV detector set at 265 nm are used. Statistical evaluation of data from participating laboratories show the average reproducibility and repeatability of the method across all samples to be excellent, with RSDR and RSDr values of 13.48 and 9.44, respectively, after elimination of outliers. The LC method for determination of vitamin D in infant formulas and enteral products has been adopted by AOAC INTERNATIONAL.

Chromatography, High Pressure Liquid↗

Intestinal absorption of copper from drinking water containing fulvic acids and an infant formula mixture studied in a suckling rat model.

The purpose of this study was to investigate if the intestinal absorption of copper in drinking water is altered in the presence of complexing agents from a fulvic acid mixture and an infant formula powder. Ten to twelve day old rat pups were given a single oral dose of radio-labeled Cu in deionized water (0.93 mg Cu/l), in water containing fulvic acids (10 mg/l), in infant formula mixed with deionized water, or in infant formula mixed with water containing fulvic acids. Six hours after dosage, radioactive Cu was analyzed in the mucosa of the small intestine, the liver and the remaining carcass (excluding the liver and gastrointestinal tract) by gamma counting. Dialysis and centrifugation experiments showed that Cu was complexed by components in the fulvic acid and formula mixtures, although the presence of fulvic acids in the water did not alter the Cu fractionation in the formula. The fractional Cu uptake (% of dose) from the intestinal lumen to the mucosa was not markedly changed by the presence of the chelating agents. However, the retention of Cu in the intestinal mucosa was increased by both fulvic acids and formula. Concomitantly, the absorption rate of Cd to the circulatory system was decreased. No interactive effect between fulvic acids and formula was found on the Cu absorption. These findings indicate that the water quality may be an important determinant of the rate of intestinal Cu absorption from drinking water. Moreover, in the future risk assessment of copper in drinking water, the possibility of alterations in absorption of drinking-water Cu has to be considered when the drinking water is used for cooking.

Animals↗

Mineral content of infant formula after treatment with sodium polystyrene sulfonate or calcium polystyrene sulfonate.

Options for the management of infants with hyperkalemia secondary to renal insufficiency are limited for infants not maintained on dialysis. Precipitation of potassium (K+) from infant formula with sodium polystyrene sulfonate (KX) prior to feeding has been reported. However, its effect on calcium (Ca2+) and sodium (Na+) has not been quantitatively defined. The purpose of this study was to examine the effects of two K+ exchange resins: KX and calcium polystyrene sulfonate (RC) on Na+, K+, and Ca2+. Infant formula powder (Similac PM 60/40, Ross Laboratories, Columbus, OH, USA) was prepared with deionized water (DW) and KX or RC (1 g/mEq of K+) was added. The formula was decanted after 50 minutes and Na+, K+, and Ca2+ were assayed in the supernatant. Na+ and K+ were also assayed in ready-to-feed PM 60/40 (RTF). KX decreased the K+ concentration by 4.5 fold (P < 0.001) and increased the Na+ content by 3.8 fold (P < 0.001). In RC there was a 1.6-fold increase in Ca2+ content (P < 0.001), and a 13% decrease in K+ concentrations (P < 0.05). Preparation of the formula with DW reduced the K+ concentration by 30% compared to ready-to-feed formula (P < 0.001). We conclude that, although KX significantly reduces the K+ content of formulas, DW may be a more practical and convenient method of preparing formula for the hyperkalemic infant.

Calcium↗

Heat treatment of infant formula: effect on postprandial serum alpha-amino-nitrogen concentrations in very-low-birth-weight infants.

Several different types of heat treatment are used in the production of commercial infant formulas. The extent of heat treatment can have different physicochemical effects on the milk proteins and will affect their solubility and digestibility. In the present study, 40 very-low-birth-weight infants were randomized at the age of 2 weeks to be given one test feed of either fresh, human milk protein-fortified human milk, a conventional canned sterilized liquid formula, a spray-dried powder formula, or an ultra-high-temperature (UHT)-treated liquid formula. The mean volume of the nasogastric test feed was adjusted so that protein intake was the same in each study group (0.45 g/kg). Venous blood samples were taken preprandially and at 30, 60, and 120 min after the test meal. alpha-amino-nitrogen in serum was assayed by the ninhydrin reaction. The concentration of alpha-amino-nitrogen rose rapidly after the fortified human milk feed, reaching a peak at 30 min. After the peak was reached, the serum values progressively declined, reaching baseline values at 120 min after the start of the test meal. The areas under the curve for all formulas were significantly different from that found for fortified human milk. At 30 min, all formulas had significantly lower alpha-amino-nitrogen values than fortified human milk; at 60 min, all formulas had significantly higher mean concentrations than fortified human milk. At 120 min, none of the formula-fed infants had reached baseline values; in particular, canned sterilized formula was still significantly above baseline values.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

Trace determination of bisphenol A and phytoestrogens in infant formula powders by gas chromatography-mass spectrometry.

This investigation describes a reliable and sensitive method for simultaneously determining bisphenol A (BPA) and two major phytoestrogens, daidzein and genistein, in powdered milks and infant formulas by gas chromatography-mass spectrometric analysis after trimethylsilylation. To reduce the matrix interference associated with the constituents of the formulas, the dissolved formula solutions were firstly ultra-centrifuged and the analytes in the supernatant were then extracted using a C18 solid-phase extraction column. The accuracy and precision of the method were determined and the technique was successfully employed to measure trace concentrations of BPA, daidzein and genistein in powdered formulas. The results show that BPA, daidzein and genistein were detected in all the testing samples (n = 6) at concentrations from 45 to 113 ng/g (except one infant formula), 20 to 2050 ng/g and 21 to 6510 ng/g, respectively. The highest concentrations of daidzein and genistein (i.e., 2050 and 6510 ng/g) were detected in a soy-based powdered infant formula. The quantitation limits were 1.0 ng/g for BPA, and 10 ng/g for daidzein and genistein using 0.5 g powdered milk samples.

Benzhydryl Compounds↗

Amino acid rating method for evaluating protein adequacy of infant formulas.

Amino acid profiles and/or protein digestibility (by the rat balance method) were determined for various forms (powder, ready-to-use, liquid concentrate, etc.) of cow's milk- and soy-based infant formulas obtained from 4 manufacturers. The essential amino acid data of the formulas were compared with that of human milk for the calculation of amino acid scores (based on the single most limiting amino acid). The product of amino acid score and total protein (g/100 kcal) was then termed "amino acid rating." Amino acid scores for the milk- and soy-based formulas ranged from 59 to 90 and from 59 to 81%, respectively, due to deficiencies in sulfur amino acids and/or tryptophan. Because of significantly higher total protein contents (g/100 kcal) of soy- (2.65-3.68) and milk-based (2.20-2.95) infant formulas compared to human milk (1.5), the relative amino acid ratings (human milk = 100) for all infant formulas except 2 liquid concentrates (having values of 87%) were above 100%. Values for true digestibility of protein in milk- and soy-based formulas ranged from 87 to 97 and from 92 to 95%, respectively. When corrected for protein digestibility, the relative amino acid ratings for all the milk-based liquid concentrates were below 100% (77-98%).

Amino Acids↗

Aluminium content of infant formulas.

An investigation was undertaken in order to assess Al concentration levels in infant formulas. Eligibility criteria were defined and strictly adhered to. An overall strategy was worked out to guarantee reliability and representativity of experimental data, in particular by minimizing all possible sources of chemical contamination or loss. The study was carried out by means of Inductively-Coupled Plasma Atomic Emission Spectrometry (ICP-AES). In cow's milk-based infant formulas as made up for consumption (about 13% in the final solution), Al levels of between 0.03-0.85 mg/L were detected. Higher amounts were present in soya-based formulas, with concentrations ranging from 0.39 to 1.01 mg/L.

Aluminum↗

An animal model of necrotizing enterocolitis induced by infant formula and ischemia in developing piglets.

BACKGROUND/AIMS: The lipid component of piglet formula (0.5% fat) causes increased mucosal permeability in 1-day-old piglets after ischemia/reperfusion. The present study examined if luminal exposure to infant formulas (3.5% fat) and ischemia/reperfusion result in an animal model of necrotizing enterocolitis and if injury is dependent on the formula fat composition. METHODS: Plasma-to-lumen clearance of 51Cr-ethylenediamine-tetraacetic acid was measured, and morphology was evaluated during luminal perfusion with preterm, term, and delipidated preterm cow milk-based infant formulas before and after ischemia/reperfusion in 1-day-old and 1-month-old piglet jejunoileum. In a separate set of experiments, a 1-2-cm segment of ileum was exteriorized and opened to expose the mucosal surface, and the villi were superfused with the above formulas (no ischemia). RESULTS: Before ischemia, clearances were markedly higher for intestinal loops perfused with preterm formula than for loops perfused with term and delipidated formulas in 1-day-old animals. After ischemia, clearances in loops perfused with preterm formula were significantly greater and grossly hemorrhagic and histologically necrotic compared with loops perfused with delipidated formula (minimal injury). Superfusion with preterm formula caused diffuse hyperemia and hemorrhage into intestinal villi. CONCLUSIONS: Luminal perfusion of 1-day-old piglet jejunoileum with predigested and bile acid-solubilized preterm infant formula, in combination with ischemia/reperfusion, produces an animal model of necrotizing enterocolitis, but only if the lipid fraction of the formula is present.

Animal Feed↗

Breast milk versus infant formulas: effects on intestinal blood flow in neonates.

The differences between breast milk and infant formulas have been a popular subject of many recent studies. Most concern the chemical and biological characteristics of breast milk and infant formulas, but little work has been done about hemodynamic changes in the splanchnic circulation. In term neonates (n = 22) we evaluated the effect of breast milk, adapted cow's milk formula, and nucleotide supplemented cow's milk formula on intestinal blood flow. To determine the blood flow velocity and estimate volume flow, pulsed Doppler ultrasound of the superior mesenteric artery (SMA) was performed prefeeding and 15, 45, and 90 minutes following feeding. When pre- and postprandial blood flow features of babies were compared among in their groups according to nutrition post prandial blood flow velocity and volume flow were increased significantly over baseline in all three groups. While there was no significant difference between the postprandial blood flow parameters of the breast milk and adapted cow's milk formula-fed groups, the nucleotide supplemented cow's milk formula-fed group had significantly higher postprandial blood flow velocity and volume flow.

Humans↗

Stability of recombinant human alpha-1-antitrypsin produced in rice in infant formula.

Human milk contains several biologically active proteins that benefit the breast-fed infant. In order to survive in the gastrointestinal tract, these proteins need to be protected against proteolysis. Since human milk contains relatively high concentrations of alpha-1-antitrypsin (AAT), we have expressed recombinant AAT in rice to explore the possibility of supplementing infant formula with this protein. The stability of recombinant AAT was examined by biochemical and functional assays, such as SDS-PAGE, Western blotting, ELISA, elastase and trypsin inhibition, following exposure to heat, low pH, and in vitro digestion, conducted in both phosphate buffered saline and infant formula. Native AAT is resistant to acidic environments down to pH 2 for 1 h and can withstand in vitro digestion modeled after conditions in the infant gut. Recombinant AAT is nearly as resistant as the native form in buffer, and is equally resilient in formula. Heat treatments (60 degrees C for 15 min, 72 degrees C for 20 sec, 85 degrees C for 3 min, and 137 degrees C for 20 sec) revealed that recombinant AAT is not as stable as native AAT in buffer, particularly at higher temperatures. While significantly less recombinant AAT is detected by ELISA after heating in formula, addition of bile extract can restore epitopes resulting in higher concentrations, suggesting protein aggregation that may not affect AAT activity. This study shows that recombinant AAT may survive the conditions of the infant stomach and duodenum and affect protein digestion in the infant small intestine.

Blotting, Western↗

Upper limits of nutrients in infant formulas: polyunsaturated fatty acids and trans fatty acids.

As upper limits for standard infant formulas, it is suggested that n-6 fatty acids not exceed 20% of total fatty acids or 10% of total energy; that alpha-linolenic acid not exceed 3% and eicosapentaenoic and docosahexaenoic acid not exceed 1% of total fatty acids; or that the sum of these n-3 fatty acids not exceed 2% of total energy. Trans fatty acids are not recommended for use in standard infant formulas. The rationale for these recommendations is presented in each case.

Fatty Acids↗

Determination of ultratrace levels of lead in infant formula by isotope dilution inductively coupled plasma mass spectrometry.

A simple method was developed for the accurate and precise determination of low- and sub-ppb (ng/g) concentrations of lead in infant formula by isotope dilution inductively coupled plasma mass spectrometry using ultrasonic nebulization. After addition of a known amount of 207Pb, samples were microwave digested and the ratio 207Pb/208Pb was measured in the digests. Agreement with certified values for lead in milk powder standard reference materials was good, and isotope dilution analysis using 206Pb yielded identical results for the standard reference materials. Lead concentrations determined for several infant nutritional products were verified by an independent method. Typically, relative standard deviations of < 4% were obtained with this method for lead concentrations above 2 ppb. The recovery of 2 ng of lead from an aqueous standard carried through the microwave digestion was 104 +/- 4%. Infant formula (containing 0.6 ppb lead) to which 0.4 ng of natural-abundance lead had been added, to simulate a formula containing 0.9 ppb lead, was analyzed by isotope dilution, and the result was 96 +/- 18% of the theoretical value. Thus, differences of 0.3 ppb lead could be clearly distinguished, and the detection limit was estimated to be 0.1 ng lead per gram of infant formula. The keys to accuracy for this method are minimizing contamination and accurately determining the concentration of lead in the isotopically enriched standard.

Food Contamination↗

[Special infant formulas].

INTRODUCTION: Breast feeding is the optimal mode of feeding for the normal fullterm infant. Formulas prepared to substitute total or partially human milk were available by the end of the last century. The composition of infant formulas has evolved greatly, because of the advance of scientific researches, and a large array of formulas for particular circumstances is actually available. OBJECTIVES: The aim of this article is to revise, compare and discuss the special manufactured formulas commercially available. MATERIAL AND METHODS: Record of the data from the manufacturer's advise leaflet or formulary for each product, and the information provided by each manufacturer after being contacted with them. RESULTS: Today there is an ample offer of specific formulas in the market. We present the composition of each specific formulas grouped as: Formulas for premature, modified formulas in its basic constituents and formulas with addition of thickeners. The general characteristics of each group are described, as well as indications and possible risks of their use. CONCLUSIONS: Knowledge of the characteristics of specific formulas is crucial for a correct prescription and choice of the most suitable one for each individual case.

Dietary Proteins↗

Anaphylactic sensitivity of guinea-pigs drinking different preparations of cows' milk and infant formulae.

Guinea-pigs have been given various preparations of cows' milk or infant formulae to drink in an investigation of the capacity of these milk preparations to stimulate per os anaphylactic sensitivity. The treatment, presumably heat, used to concentrate the 'Evaporated' whole cows' milks (three brands were tested) almost abolished their sensitizing capacity to beta lactoglobulin. The brand presumed to be most heated as judged by the degree of caramelization had also lost most of its sensitizing capacity to casein. Injected parenterally, the 'Evaporated' milk adequately sensitized to anaphylaxis. An infant formula, which in its spray dried form was only moderately sensitizing to both beta lactoglobulin and casein, lost most of this sensitizing capacity when processed to a liquid concentrate state. The manufacturing process for the liquid concentrate involved greater and more prolonged heating than required for the spray-dried form. The relevance of these findings in the guinea-pig to cows' milk protein intolerance in the infant and possibly also to cot death is discussed.

Anaphylaxis↗

Prebiotic effect of fructo-oligosaccharide supplemented term infant formula at two concentrations compared with unsupplemented formula and human milk.

BACKGROUND: Human milk components, including oligosaccharides, affect the gastrointestinal flora of infants. Previous studies in adults have demonstrated that fructo-oligosaccharides increase potentially beneficial fecal bacteria, including bifidobacteria. The purpose of this study was to determine the prebiotic effect of infant formula supplemented with fructo-oligosaccharides. METHODS: Healthy term infants 2 to 6 weeks of age were enrolled in a 5-week, prospective, randomized, crossover, single-site study with a nonrandomized human milk comparator group. Washout weeks preceded and followed a week of feeding with fructo-oligosaccharide-supplemented formula (1.5 or 3.0 g/L). Stool specimens were quantitatively cultured weekly for bacteroides, lactobacilli, bifidobacteria, clostridia and enterococci and were tested for Clostridium difficile toxin. RESULTS: Seventy-two of 87 infants completed the trial; 58 were formula fed and 14 were human milk fed. Mean counts of bifidobacteria and lactobacilli were similar in all groups at entry and no group experienced a significant change in counts with fructo-oligosaccharide supplementation. After 7 days of fructo-oligosaccharide supplementation the bifidobacteria counts were greater in the 1.5 g/L fructo-oligosaccharide formula group than in the human milk fed or 3.0 g/L fructo-oligosaccharide formula groups. Formula-fed infants had higher counts of enterococci and bacteroides before fructo-oligosaccharide supplementation, and these counts did not change after supplementation. Clostridium counts increased 7 days after supplementation in the 1.5 g/L fructo-oligosaccharide formula group (P = 0.0356). No human milk fed infants had C. difficile toxin in stools. Fructo-oligosaccharide (3.0 g/L) supplementation resulted in more frequent and significantly softer stools. CONCLUSIONS: Infant formula supplemented with 1.5 or 3.0 g/L fructo-oligosaccharides was safe but had minimal effect on fecal flora and C. difficile toxin.

Bifidobacterium↗

Availabilities of calcium, iron, and zinc from dairy infant formulas is affected by soluble dietary fibers and modified starch fractions.

OBJECTIVE: Insoluble dietary fiber is a known inhibitor of mineral absorption, whereas the effects of soluble dietary fibers (including prebiotics) are less known. The aim was to study calcium, iron, and zinc availabilities from dairy infant formulas supplemented with soluble dietary fibers and modified starches in vitro. METHODS: Dairy infant formulas were supplemented with soluble dietary fibers (3%, dry wt) and modified starches (16% pregelatinized rice starch and 1.9% maltodextrin, dry wt) and kept in a well-controlled and defined environment in vitro. Pooled mature human milk was used as the reference standard. RESULTS: Calcium availability from standard formula was elevated by 30% after inulin supplementation (17.2%), whereas locust bean gum (11.9%) and high esterified pectin (11.7%) reduced availability by approximately 10%. Iron availability from standard formula was increased by pregelatinized rice starch (3.8%), whereas availability was reduced in the following order: high esterified pectin (2.3%), oligofructose (2.2%), and low esterified pectin (2.1%). Zinc availability was highest after the addition of pregelatinized rice starch (13.5%) but lowest with the addition of locust bean gum (6.8%) and maltodextrin (5.4%). CONCLUSIONS: This study showed that addition of soluble dietary fiber affects calcium, iron, and zinc availabilities in positive (inulin) and negative ways, depending on the type of the dietary fiber used.

Biological Availability↗